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Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells
Matrix attachment regions (MARs) can mediate the replication of vector episomes in mammalian cells; however, the molecular mode of action remains unclear. Here, we assessed the characteristics of MARs and the mechanism that mediates episomal vector replication in mammalian cells. Five shortened subf...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6789156/ https://www.ncbi.nlm.nih.gov/pubmed/31509492 http://dx.doi.org/10.1091/mbc.E19-02-0108 |
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author | Wang, Xiao-Yin Zhang, Xi Wang, Tian-Yun Jia, Yan-Long Xu, Dan-Hua Yi, Dan-Dan |
author_facet | Wang, Xiao-Yin Zhang, Xi Wang, Tian-Yun Jia, Yan-Long Xu, Dan-Hua Yi, Dan-Dan |
author_sort | Wang, Xiao-Yin |
collection | PubMed |
description | Matrix attachment regions (MARs) can mediate the replication of vector episomes in mammalian cells; however, the molecular mode of action remains unclear. Here, we assessed the characteristics of MARs and the mechanism that mediates episomal vector replication in mammalian cells. Five shortened subfragments of β-interferon MAR fragments were cloned and transferred into CHO cells, and transgene expression levels, presence of the gene, and the episomal maintenance mechanism were determined. Three shortened MAR derivatives (position 781–1320, 1201–1740, and 1621–2201) retained full MAR activity and mediated episomal vector replication. Moreover, the three shortened MARs showed higher transgene expression levels, greater efficiency in colony formation, and more persistent transgene expression compared with those of the original pEPI-1 plasmid, and three functional truncated MARs can bind to SAF-A MAR-binding protein. These results suggest that shortened MARs are sufficient for replication and maintenance of episomes in CHO cells. |
format | Online Article Text |
id | pubmed-6789156 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-67891562019-12-30 Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells Wang, Xiao-Yin Zhang, Xi Wang, Tian-Yun Jia, Yan-Long Xu, Dan-Hua Yi, Dan-Dan Mol Biol Cell Article Matrix attachment regions (MARs) can mediate the replication of vector episomes in mammalian cells; however, the molecular mode of action remains unclear. Here, we assessed the characteristics of MARs and the mechanism that mediates episomal vector replication in mammalian cells. Five shortened subfragments of β-interferon MAR fragments were cloned and transferred into CHO cells, and transgene expression levels, presence of the gene, and the episomal maintenance mechanism were determined. Three shortened MAR derivatives (position 781–1320, 1201–1740, and 1621–2201) retained full MAR activity and mediated episomal vector replication. Moreover, the three shortened MARs showed higher transgene expression levels, greater efficiency in colony formation, and more persistent transgene expression compared with those of the original pEPI-1 plasmid, and three functional truncated MARs can bind to SAF-A MAR-binding protein. These results suggest that shortened MARs are sufficient for replication and maintenance of episomes in CHO cells. The American Society for Cell Biology 2019-10-15 /pmc/articles/PMC6789156/ /pubmed/31509492 http://dx.doi.org/10.1091/mbc.E19-02-0108 Text en © 2019 Wang, Zhang, et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0 This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License. |
spellingShingle | Article Wang, Xiao-Yin Zhang, Xi Wang, Tian-Yun Jia, Yan-Long Xu, Dan-Hua Yi, Dan-Dan Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells |
title | Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells |
title_full | Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells |
title_fullStr | Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells |
title_full_unstemmed | Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells |
title_short | Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells |
title_sort | shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6789156/ https://www.ncbi.nlm.nih.gov/pubmed/31509492 http://dx.doi.org/10.1091/mbc.E19-02-0108 |
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